Objective To investigate the regulatory function of ubiquitin-like modifier activating enzyme 1 (Uba1) in the growth and development of myoblasts.

Methods C2C12 myoblasts were used as a model to establish cell models with Uba1 overexpression and knockdown. Myogenic differentiation was assessed by immunofluorescence staining for myosin heavy chain (MyHC). Cell migration was analyzed using a wound-healing assay, and cell morphological changes were observed through cell adhesion and spreading assays. The expression of differentiation and migration marker genes was further examined by qRT-PCR and Western blotting.

Results C2C12 cell models with stable Uba1 overexpression and knockdown were successfully generated. Overexpression of Uba1 significantly increased the number of MyHC-positive cells on day 5 of differentiation and markedly upregulated the expression of the myogenic differentiation markers MyoG and MyHC at both the mRNA and protein levels. It also promoted cell migration, with a significantly increased migration distance 24 h after scratching and upregulated the expression of migration-related genes Rac1, Fak, and Paxillin. Furthermore, Uba1 overexpression significantly increased the number of adherent cells and markedly increased the average cell spreading area. Conversely, knockdown of Uba1 produced the opposite effects on all the above parameters.

Conclusion Uba1 promotes the differentiation, migration, adhesion, and spreading of C2C12 cells, thereby exerting a positive regulatory role in the growth and development of skeletal muscle.